Measurement of S-adenosylmethionine and S-adenosylhomocysteine in mice liver with high performance liquid chromatography
Objective To develop a method for determination of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) simultaneously in mice liver by reversed-phase high performance liquid chromatography(HPLC).Methods Mice livers were weighed,emulsified quickly by handheld homogenate,four times the volume of 0.6 mol/L HCLO4added for ultrasonic,and the hydrolysate was adjusted to centrifuged and filtrated with a membrane.The supernatants were separatedonVenusil MP-C18 column (250 mm ×4.6 mm,5 μm) at 30 ℃ with a mobile phase of 50 mmol/L NaH2PO4,10 mmol/L C7H15NaO3S and methanol,a flow rate of 1.0 ml/min and UV detection at 254 nm.Results The SAM and SAH in the corresponding concentration range showed a good linear relation with its peak area,correlation coefficient (r ＞ 0.9990),recovery was 92.20％-101.38％,RSD was 2.88％-6.78％.The average within-day precision of SAM and SAH was 4.14％ and 3.71％,and the average day to day precision was 7.51％ and 9.54％.The content of SAM and SAH in mice liver was 3.14-6.09 mg/L (31.44 -60.98 nmol/g wet weight) and 1.29-3.10 mg/L (13.38-32.17 nmol/g wet weight) respectively.Conclusion The validated method is simple,rapid accurate and reliable to the determination of SAM and SAH in mice liver.
NETL
NSTL
万方数据
维普
